中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
5期
818-820
,共3页
逯翀%刘春萍%郭辉%田元%张景辉
逯翀%劉春萍%郭輝%田元%張景輝
록충%류춘평%곽휘%전원%장경휘
乳腺肿瘤%干细胞%Notch1
乳腺腫瘤%榦細胞%Notch1
유선종류%간세포%Notch1
Breast neoplasm%Stem cell%Notch1
目的 检测乳腺癌细胞株MCF-7与MCF-7/ADR的 Notch1 表达、成球能力、干细胞池比例及干细胞内Notch1的表达,探讨Notch1 在逆转乳腺癌化疗耐药中的意义.方法 实时荧光定量逆转录-聚合酶链反应(RT-qPCR)及Western blot 法检测乳腺癌细胞株MCF-7、MCF-7/ADR 及表型为ALDH1+/CD44+/CD24- 乳腺癌干细胞中Notch1 的表达；无血清培养检测MCF-7及MCF-7/ADR 的成球能力；流式细胞仪检测MCF-7与MCF-7/ADR的干细胞比例并分选干细胞.结果 MCF-7/ADR的Notch1 的表达明显高于MCF-7(0.0748±0.0043比0.0461±0.0022,P＜0.05),微球体形成能力也明显高于MCF-7 (P＜0.05)；流式细胞仪检测干细胞比例,MCF-7/ADR高于MCF-7(11.40％比1.89％)；分选出的干细胞Notch1 的表达明显高于非干细胞(0.3096±0.0324比0.0428±0.0061,P ＜0.05).结论 Notch1 在乳腺癌及乳腺癌干细胞中高表达,干预Notch1 可能可以逆转乳腺癌治疗的耐药.
目的 檢測乳腺癌細胞株MCF-7與MCF-7/ADR的 Notch1 錶達、成毬能力、榦細胞池比例及榦細胞內Notch1的錶達,探討Notch1 在逆轉乳腺癌化療耐藥中的意義.方法 實時熒光定量逆轉錄-聚閤酶鏈反應(RT-qPCR)及Western blot 法檢測乳腺癌細胞株MCF-7、MCF-7/ADR 及錶型為ALDH1+/CD44+/CD24- 乳腺癌榦細胞中Notch1 的錶達；無血清培養檢測MCF-7及MCF-7/ADR 的成毬能力；流式細胞儀檢測MCF-7與MCF-7/ADR的榦細胞比例併分選榦細胞.結果 MCF-7/ADR的Notch1 的錶達明顯高于MCF-7(0.0748±0.0043比0.0461±0.0022,P＜0.05),微毬體形成能力也明顯高于MCF-7 (P＜0.05)；流式細胞儀檢測榦細胞比例,MCF-7/ADR高于MCF-7(11.40％比1.89％)；分選齣的榦細胞Notch1 的錶達明顯高于非榦細胞(0.3096±0.0324比0.0428±0.0061,P ＜0.05).結論 Notch1 在乳腺癌及乳腺癌榦細胞中高錶達,榦預Notch1 可能可以逆轉乳腺癌治療的耐藥.
목적 검측유선암세포주MCF-7여MCF-7/ADR적 Notch1 표체、성구능력、간세포지비례급간세포내Notch1적표체,탐토Notch1 재역전유선암화료내약중적의의.방법 실시형광정량역전록-취합매련반응(RT-qPCR)급Western blot 법검측유선암세포주MCF-7、MCF-7/ADR 급표형위ALDH1+/CD44+/CD24- 유선암간세포중Notch1 적표체；무혈청배양검측MCF-7급MCF-7/ADR 적성구능력；류식세포의검측MCF-7여MCF-7/ADR적간세포비례병분선간세포.결과 MCF-7/ADR적Notch1 적표체명현고우MCF-7(0.0748±0.0043비0.0461±0.0022,P＜0.05),미구체형성능력야명현고우MCF-7 (P＜0.05)；류식세포의검측간세포비례,MCF-7/ADR고우MCF-7(11.40％비1.89％)；분선출적간세포Notch1 적표체명현고우비간세포(0.3096±0.0324비0.0428±0.0061,P ＜0.05).결론 Notch1 재유선암급유선암간세포중고표체,간예Notch1 가능가이역전유선암치료적내약.
Objective To investigate the role of Notch1 in the chemotherapy process of breast cancer by studying the the efficiency of mammosphere formation,the ratio of stem cells in MCF-7 and MCF-7/ADR,and the expression status of Notch1 in MCF-7,MCF-7/ADR and stem cells respectively.Methods Real time fluorescent quantitative reverse transcriptase polymerase chain reaction (RT-qPCR)and Western blotting were employed to detect the expression status of Notch1 in breast cancer cell lines MCF-7,MCF-7/ADR and ALDH1 +/CD44 +/CD24 - breast cancer stem cells (BCSCs).Serum-free medium was available to assess the efficiency of mammospbere formation.Flow cytometry was used to define the ratio of BCSCs and select them in MCF-7 and MCF-7/ADR.Results The expression status of Notehl [ (0.0748 ± 0.0043 ) vs ( 0.0461 ± 0.0022 ),P ＜ 0.05 ] and the ability of mammosphere formation were significantly higher in MCF-7/ADR than in MCF-7 ( P＜0.05).There was significant difference in the ratio of BCSCs between MCF-7/ADR and MCF-7 ( 11.40％ vs 1.89％ ).Furthermore the expression score of Notch1 in BCSCs was significantly higher than in non-BCSCs [ (0.3096 ± 0.0324 ) vs (0.0428 ±0.0061 ),P ＜0.05].Conclusion The high level expression of Notch1 in breast cancer and BCSCs suggests regulating Notchl may play an important role in reversing chemotherapy resistance of breast cancer.
